盐酸青藤碱诱导人肝癌细胞SMMC-7721凋亡及其死亡受体DR4、DR5表达的变化

Sinomenine hydrochloride inducing apoptosis of hepatocellular carcinoma SMMC-7721 cells and its effect on expressions of DR4 and DR5

目的观察人肝癌细胞SMMC-7721凋亡过程中死亡受体DR4、DR5的表达,探讨盐酸青藤碱诱导人肝癌细胞凋亡的作用机制。方法体外培养人肝癌细胞SMMC-7721,经不同浓度盐酸青藤碱作用后,采用MTT法检测细胞增殖情况;Hoechst染色荧光显微镜观察细胞凋亡;用Western blotting方法检测人肝癌细胞DR4、DR5的表达。结果盐酸青藤碱对人肝癌细胞的生长有明显的抑制作用,并呈剂量、时间依赖性,不同浓度组之间差异均有统计学意义(P<0.05);Hoechst33258染色可见凋亡细胞皱缩,核碎裂,也可见典型凋亡小体;在蛋白水平,盐酸青藤碱处理后SMMC-7721细胞死亡受体DR4、DR5的表达水平较未处理组显著增加(P<0.05)。结论盐酸青藤碱可抑制人肝癌细胞SMMC-7721的细胞增殖,促进其凋亡,此作用可能与细胞内死亡受体DR4和DR5表达上调有关。

Objective To investigate the expression levels of DR4 and DR5 on the process of apoptosis induced by Sinomenine hydrochloride in hepatocellular carcinoma（ HCC） SMMC-7721 cells. Methods MTT assay was used to determine the effect of Sinomenine hydrochloride on the growth of hepatocellular carcinoma SMMC-7721 cells. In vitro cultured human HCC SMMC-7721 cells： the effect of Sinomenine hydrochloride on the morphologic change induced by sinomenine hydrochloride were assessed by fluorescence microscope after Hoechst staining. SMMC-7721 cells were cultured for 48 hours with Sinomenine hydrochloride 2 mmol / L,then the effect of Sinomenine hydrochloride on death receptors（ DR4 and DR5） were detected by Western blotting. Results Sinomenine hydrochloride inhibited the growth of human HCC SMMC-7721 cells in a concentration-and time-dependent manner as determined by MTT assay. In vitro cultured human HCC SMMC-7721 cells,Sinomenine hydrochloride induced apoptosis by Hoechst staining; Sinomenine hydrochloride significantly increased death receptors（ DR4 and DR5） protein expression in a concentration-dependent manner in human HCC SMMC-7721 cells. Conclusion Sinomenine hydrochloride can inhibit growth of human HCC SMMC-7721 cells through apoptotic induction; Sinomenine hydrochloride can increase the expression levels of death receptors（ DR4 and DR5） on the process of apoptosis induced by Sinomenine hydrochloride in HCC SMMC-7721 cells.